CN108663517A - It can be used for diagnosing the kit of human enterovirus infection - Google Patents
It can be used for diagnosing the kit of human enterovirus infection Download PDFInfo
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- CN108663517A CN108663517A CN201710208431.0A CN201710208431A CN108663517A CN 108663517 A CN108663517 A CN 108663517A CN 201710208431 A CN201710208431 A CN 201710208431A CN 108663517 A CN108663517 A CN 108663517A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56983—Viruses
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/005—Assays involving biological materials from specific organisms or of a specific nature from viruses
- G01N2333/08—RNA viruses
- G01N2333/085—Picornaviridae, e.g. coxsackie virus, echovirus, enterovirus
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2469/00—Immunoassays for the detection of microorganisms
- G01N2469/20—Detection of antibodies in sample from host which are directed against antigens from microorganisms
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Abstract
The present invention provides the kit that can be used for diagnosing human enterovirus infection.The detection kit of the present invention includes one or more solid carriers and the SEQ ID NO being independently connected on the solid carrier：Polypeptide shown in 1~6.When with whether SEQ ID NO：Whether polypeptide shown in 1~6 all has response as judgement subject in the case of index infect by human enterovirus the IgM in the blood sample in subject source, can diagnose human enterovirus with 92% or more sensitivity and 6% false positive rate below and infect.
Description
In the complete process of the present invention, cooperation unit Shanghai Public Health Clinical Center, Suzhou Yvonne consor have been obtained
The support energetically of object Materials Co., Ltd indicates sincere and thanks herein.
Technical field
The invention mainly relates to detection kits.Specifically, the present invention relates to can be used for detecting human enterovirus infection
Kit.
Background technology
Enterovirus (EV) infection is the disease caused by human enterovirus, is clinically mainly shown as such as brothers mouthful
Disease, hand-foot-and-mouth disease eruption and prevalence mostly in infant, part EV infection infants show as herpangina, and patient with severe symptoms goes out
Existing viral encephalitis, viral cerebrospinal meningitis, pulmonary edema, empsyxis etc..Cause the cause of disease of hand-foot-and-mouth disease, most commonly
Coxsackie virus A 6 and enterovirns type 71；The large percentage of severe infection, disease occur for the disease caused by enterovirus infection
Dead rate is also higher, and severe cases case fatality rate is up to 10%-25%, therefore etiological diagnosis is significant.
Three aspects of clinical diagnosis Main Basiss of enterovirus:When clinical manifestation, second is that serological evidence, third, disease
Original learns evidence.Since enterovirus EV infection clinical manifestations are more, the Serologic detection and pathogeny detection of enterovirus are
Current most important foundation.EV infection generates immunoprotection, and IgM and IgG antibody can be detected after infection.
Invention content
The purpose of the present invention is to provide the new kits and diagnostic method that can be used for diagnosing human enterovirus infection.
The present inventor it has surprisingly been found that when with whether SEQ ID NO：Polypeptide shown in 1~6 all carrys out subject
IgM in the blood sample in source has response as judging subject whether in the case of the index infected by human enterovirus,
It can be with 92% or more sensitivity and 6% false positive rate diagnosis human enterovirus infection below.
Therefore, the present invention includes：
1. a kind of detection kit comprising one or more solid carriers, and be independently connected to the solid and carry
SEQ ID NO on body：Polypeptide shown in 1~6.
2. according to the detection kit described in item 1, wherein the SEQ ID NO：Polypeptide shown in 1~6 is independently connected
In on same solid carrier.
3. according to the detection kit described in item 1, wherein the solid carrier is SJ modified silica-gels.
4. according to the detection kit described in item 1, it is used to diagnose human enterovirus infection.
5. a kind of diagnosis subject whether be people's enterovirus infection method, this method includes：
The SEQ ID NO are detected using the detection kit described in any one of item 1~4：Polypeptide pair shown in 1~6
Whether the IgM in the blood sample in subject source has response；Wherein,
As the SEQ ID NO：Polypeptide shown in 1~6 all has response to the IgM in the blood sample in subject source
When, judgement should subject is a human enterovirus infection；Otherwise, it is determined that the subject is inhuman enterovirus infection.
6. according to the method described in item 5, wherein the blood sample is whole blood, blood plasma or serum.
7. according to the method described in item 5, wherein the subject is the subject with hand-foot-and-mouth disease clinical symptoms.
The specific implementation mode of invention
Polypeptide
SEQ ID NO：Polypeptide sequence shown in 1 is YHSSVYSLPPDPDHFDGYKQ；
SEQ ID NO：Polypeptide sequence shown in 2 is HPYVLDAGIPISQLTVCPHQ；
SEQ ID NO：Polypeptide sequence shown in 3 is ISDLLASVDSEEVRQYCRDQ；
SEQ ID NO：Polypeptide sequence shown in 4 is GHFTMLGVRDRLAVLPRHSQ；
SEQ ID NO：Polypeptide sequence shown in 5 is AREKVEFLNNLKQLPLLENQ；
SEQ ID NO：Polypeptide sequence shown in 6 is AAAQKNFTMKLCKDASDILQ.
The polypeptide suitable can be obtained using the known methods such as (1) chemical synthesis process or (2) enzyme reaction synthetic method
, wherein chemical synthesis is more easy.In the polypeptide of the chemical synthesis present invention, by using peptide synthesizer synthesis or
Person semi-synthetic polypeptide carries out.As chemical synthesis process, can enumerate such as peptide solid-phase synthesis.It synthesizes in this way
Peptide conventional means may be used purified such as ion-exchange chromatography, reverse phase high performance liquid chromatography, affinity chromatography.This
The peptide solid phase synthesis process of sample with and subsequent peptide purification be all well-known in the art.
In addition, by the polypeptide of the enzyme reaction production present invention, such as International Publication pamphlet may be used
Method described in No. WO2004/011653.I.e., it is possible to produce in this way：By the amino acid of a side or the carboxyl terminal quilt of dipeptides
Amino acid or dipeptides obtained from esterification or amidation, amino acid (such as the carboxy protective for being in amino acid free state
Amino acid) it is reacted in the presence of peptide synthetase, the dipeptides or tripeptides of generation.As peptide synthetase, can enumerate：
The culture of microorganism with the ability for generating peptide, the thalline of the microbial cells or the microorganism that are detached by the culture
Processed material or the microbe-derived peptide synthetase.
The chemical synthesis of especially polypeptide has been realized in commercialization, can by entrust profession Peptide systhesis company come
Synthesize the polypeptide.
Kit
The present invention provides a kind of detection kit (kit of the invention) comprising one or more solid carriers, with
And it is independently connected to the SEQ ID NO on the solid carrier：Polypeptide shown in 1~6.
Preferably, the present invention also provides a kind of detection devices and the detection device in the kit for preparing the present invention
Purposes, the detection device includes one or more solid carriers, and is independently connected on the solid carrier
SEQ ID NO：Polypeptide shown in 1~6.
It is surprising that the kit of the present invention can be with 92% or more sensitivity and 6% false positive rate below
Diagnose human enterovirus infection.In this specification, sensitivity refers to：In positive sample with the confirmation of " goldstandard " method, by it
His method is measured as the ratio of positive sample.False positive rate refers to：In negative sample with the confirmation of " goldstandard " method, by it
His method is measured as the ratio of positive sample." goldstandard " method that the art detects human enterovirus infection is nucleic acid
Detection method, i.e. PCR methods.In this specification, " diagnosis " can be made a definite diagnosis, and can also be to provide reference information to make a definite diagnosis.
The so-called enterovirus infection of this specification, including but not limited to enterovirns type 71 infect.
In the present invention, solid carrier is not particularly limited, as long as (being, for example, can as solid or insoluble material
Material to be detached from reaction mixture by filtering, precipitation, Magnetic Isolation etc.) carrier.
Constitute solid carrier material include but not limited to：Silica gel (dimethyl silicone polymer, PDMS), cellulose, Teflon
It is grandTM, NC Nitroncellulose, agarose, glucan, chitosan, polystyrene, polyacrylamide, polyester, makrolon, polyamides
It is amine, polypropylene, nylon, polyvinylidene fluoride, latex, silica, glass, glass fibre, gold, platinum, silver, copper, iron, stainless
Steel, ferrite, silicon wafer, polyethylene, polyethyleneimine, polylactic acid, resin, polysaccharide, albumen (albumin etc.), carbon or it
Combination etc..
The shape of solid carrier includes but not be limited to：Pearl, magnetic bead, film, microcapillary, filter membrane, plate, micro plate, carbon
Nanotube, sensor chip etc..Just as known in the art, it can be set on the flat solid carrier such as film or plate
Set pit, groove, filter membrane bottom etc..
Magnetic bead can be with the sphere diameter of about 25nm~about 1mm ranges.In a preferred embodiment, magnetic bead has about
The diameter of the μ ms of 50nm~about 10.The size of magnetic bead can be selected according to specific purposes.By Sepharose etc.
Pearl made of high crosslinked spherical agarose has the diameter of about 24 μm~about 165 μ ms.Preferably, high crosslinked spherical fine jade
Lipolysaccharide pearl has the diameter of about 24 μm~about 44 μ ms.The size of high crosslinked spherical sepharose 4B can be according to specific purposes
To be selected.
The example of solid carrier with hydrophobic surface include can from Polysciences, Warrington, PA or
The polystyrene latex beads such as the product of Spherotech, Liberville, IL purchase.
Silica (SiO2)-processing or silica (SiO2) base solid carrier example include can be from
The extraordinary magnetic silica pearl etc. of Polysciences, Warrington, PA purchase, can be used for capturing nucleic acid (such as
DNA).It can be from Dynal Biotech M-280 bought etc. alternatively, can also use.
Magnetic bead with hydrophilic surface can be used for capturing bacterial cell, nucleic acid and the other ingredients of proliferation period.As
The example of the magnetic bead can enumerate Polysciences, the pearl (title of Warrington, PA sale：Biomag (registrations
Trade mark) carboxyl) or Bangs Laboratory, Inc., Fishers, IN entitled MC02N/2928 pearl.Or
Person, the M-270 etc. that Dynal Biotech can be used to sell.
In a preferred embodiment, the solid carrier is SJ modified silica-gels, and being the poly- biomaterials of Suzhou Yvonne has
A kind of microarray solid support material (the iPDMS films, referring to Chinese patent of silicone rubber material of limit company exploitation
CN101265329A).This material is that specific initiator is added wherein based on the common PDMS of biological study
Ingredient (makes the material that can realize surface-functionalized modification by surface initiated polymerization (SIP)), using polyethylene glycol first
What base acrylate (poly (oligo (ethylene glycol) methacrylate), pOEGMA) surface modification obtained.SJ
Modified silica-gel has outstanding anti-protein non-specific adsorption (Nonspecific protein adsorption, NPA) energy
Power, non-specific protein absorption control in complicated protein immunization being detected to close to " absolute 0 " it is horizontal (it is close or
Less than the detectable limit of instrument), the trouble that can not only exempt closing and be cleaned multiple times can also be by using stronger letter
Number amplification means improve the sensitivity of protein microarray.And the essence of its silicon rubber imparts the stronger machine of the material
Tool performance and good operability.SJ modified silica-gels are successfully applied to 11 tumor-markers by the poly- companies of Suzhou Yvonne
The combination assay microarray ELISA kit of object composition, realize high-throughput and highly sensitive detection, it was demonstrated that this material
Material is a kind of outstanding protein microarray solid support material.Meanwhile this material also has the adjustable spy of surface nature
Property, its surface topography can be adjusted in a certain range by the controlled modification reaction time.
The connection well known to a person skilled in the art polypeptide and solid carrier may be used in the connection of polypeptide and solid carrier
Method carries out.For example, for the connection of protein/polypeptide and modified silica-gel surface, 1- ethyl -3- (3- can be passed through
Dimethyl aminopropyl)-carbodiimides [1-ethyl-3- (3-dimethyl ami-nopropyl) carbodiimide,
EDC] and n-hydroxysuccinimide (N-hydroxysuccinimide, NHS) reaction by modified silica-gel surface macromolecule
Carboxyl (- COOH) group on chain is changed to activated group, which can be with the amino (- NH2) of institute's band on protein/polypeptide
Protein/polypeptide is fixed on solid carrier surface by reaction to realize.
The concentration of polypeptide in the sampling liquid that is used when point sample is not particularly limited, those skilled in the art can be according to normal
Rule selection, preferably 1 μ of μ g~1000 g/mL, the more preferable 10 μ μ of g~500 g/mL.In addition, dividing on a solid carrier for polypeptide
The density of cloth is not particularly limited, and those skilled in the art can be according to conventional selection, preferably 1~100 point/10mm2, more excellent
Select 5~50 points/10mm2。
In the present specification, " independent " refers to：The polypeptide forms a point on the solid carrier, is not wrapped in the point
Preferably only include the polypeptide containing on other polypeptides of the influential amount of detection.
Solid carrier in the kit of the present invention can be one, can also be multiple, for example, more than two but excellent
It is selected as 1.
The kit can also include：
1. the serum dilution or serum dilution component solution that prepare：Serum dilution, such as there is Beijing to match biology of speeding
The Sample dilution (product identification 070021-S2) of Science and Technology Ltd., the sample-adding of the Zhengzhou bio tech ltd Bo Weijia
Change colour Sample dilution (product identification bwj010103) etc..The serum dilution is used for dilute serum, the blood of kit detection
Suitable multiple, such as 2~200 times, preferably 10~100 times are diluted clearly.
The kit can also include：
2. concentrating washing lotion：After solid carrier surface is incubated serum and ELIAS secondary antibody, solid carrier table need to be washed with washing lotion
Face unbonded antibody and ELIAS secondary antibody.Concentration washing lotion is, for example, 1% polysorbas20 aqueous solution, and when use need to dilute 2~40
Again, preferably 5~20 times.
The kit can also include：
3. ELIAS secondary antibody solution：IgM in human enterovirus infected person anteserum and solid carrier (such as SJ modified silica-gels)
On polypeptide combine, ELIAS secondary antibody can be combined with IgM, and the marker on ELIAS secondary antibody can be reacted with assay chromogenic substrate solution TMB,
Generate macroscopic royal purple chrominance signal.ELIAS secondary antibody can be the goat-anti people IgM of such as horseradish peroxidase-labeled.It is right
Concentration of the ELIAS secondary antibody in ELIAS secondary antibody solution is not particularly limited, and can be such as 1ng~1000ng/mL.
4. chemical colour reaction substrate solution TMB：Chemical colour reaction substrate TMB can be with the horseradish peroxidase that is marked on ELIAS secondary antibody
Reaction, generates macroscopic royal purple chrominance signal.
The kit can also include：
5. one or more reaction cavity.
Such as can be biological incubation reactor more than one or two, it may be used according to demand for example two-sided
Biological incubation reactor, referring to Chinese patent ZL 201120177686.3 or ZL201110142518.5；Or single side
Biological incubation reactor, referring to Chinese patent ZL201220430886.x.
The kit can also include：
6. other are used to detect the detection molecules (such as polypeptide, protein, nucleic acid etc.) of human enterovirus infection.
The kit can also include：
7. operation instructions.
The kit can also include：
8. the positive quality control point being connected on one or more of solid carriers or other independent solid carriers (can
Using e.g. people IgM as positive quality control point) and/or negative Quality Control point (such as phosphate buffer, abbreviation PB).
Preferably, it can be with hand-foot-and-mouth disease clinical symptoms that kit of the invention, which can be adapted for the subject,
Subject
Diagnostic method
In another aspect, the present invention provide it is a kind of diagnosis subject whether be people's enterovirus infection method (this
The diagnostic method of invention), this method includes：
The SEQ ID NO are detected using the detection kit of aforementioned present invention：Polypeptide shown in 1~6 carrys out subject
Whether the IgM in the blood sample in source has response；Wherein,
As the SEQ ID NO：Polypeptide shown in 1~6 all has response to the IgM in the blood sample in subject source
When (criterion 1), judgement should subject is a human enterovirus infection；Otherwise, it is determined that the subject is inhuman enterovirus infection.
In the present specification, " response " refer to：Polypeptide site shows the royal purple chrominance signal stronger than negative control point.
In this specification, the positive reaction to the kit of the present invention refers to：On the blood sample in subject source meets
State criterion 1；Otherwise it is negative reaction.
The blood sample can be whole blood, blood plasma or serum.
In one preferred embodiment, kit of the invention, which can be adapted for diagnosis, has hand-foot-and-mouth disease clinical condition
Whether the subject of shape is people's enterovirus infection.In the present invention, hand-foot-and-mouth disease clinical symptoms be embodied in fever and
There are fash or bleb in the positions such as hand, foot, oral cavity.
Embodiment
1. the preparation and confirmation of polypeptide
The polypeptide used in embodiment is synthesized by gill biochemistry (Shanghai) Co., Ltd., and has been carried out really to it by mass spectrum
Recognize.Wherein,
SEQ ID NO：1YHSSVYSLPPDPDHFDGYKQ.
SEQ ID NO：2HPYVLDAGIPISQLTVCPHQ.
SEQ ID NO：3ISDLLASVDSEEVRQYCRDQ.
SEQ ID NO：4GHFTMLGVRDRLAVLPRHSQ.
SEQ ID NO：5AREKVEFLNNLKQLPLLENQ.
SEQ ID NO：6AAAQKNFTMKLCKDASDILQ.
2. the preparation of kit
Detection chip be with SJ modified silica-gels (iPDMS films) be solid support material, on it by point sample fix it is more
Peptide solution is prepared.SJ modified silica-gels are that with olefin-terminal, surface is added in traditional polydimethyl siloxane material
The initiator of initiated polymerization, and pass through three-dimensional structure of the heat cross-linking (si-h bond bonding) fixed to dimethyl silicone polymer
In, obtain a kind of new material i.e. SJ modified silica-gels.Its manufacturing process is referring to International Patent Publication WO2014/044184.
The SJ modified silica-gel films made can save in 4 DEG C of refrigerators.
Using16 people's point sample instruments of PersonalArrayerTM prepare polypeptide microarrays (i.e. on modified silica-gel
Detection chip), process is：
1) it pre-processes
By SJ modified silica-gel thin slices (15 × 15mm2) be immersed in activating solution, taking-up deionized water elution 3 after 30min
It is secondary, it is dried up with nitrogen, is immediately used to point sample.
2) point sample
Sampling liquid is diluted good and is transferred in the corresponding micropore of 384 orifice plates, 384 orifice plates with sample are placed in point sample instrument
On base station, while pretreated modified silica-gel thin slice being placed on the base station of point sample instrument, carries out point sample at once.Point sample environment item
Part is room temperature (25 DEG C), humidity set 50%.By SEQ ID NO：The point on SJ modified silica-gel thin slices of polypeptide shown in 1~6
At microarray, one point of each polypeptide point sample, in addition one people IgM point of point sample is as positive quality control point and a PB points work
For negative Quality Control point.The point sample amount each put on manufactured polypeptide microarrays is about 0.6nL, and sampling point radius is 200 μm.
3) chemistry is fixed
The polypeptide microarrays just made will be placed in climatic chamber (26 DEG C, 60% humidity) fixed at least 6h.Chemistry is solid
Process is determined referring to International Patent Publication WO2014/044184.
It will include first to capture the buffer solution point of peptide molecule on modified silica-gel film by point sample instrument, then buffer
Liquid starts to evaporate, and capture peptide molecule and the surface intimate contact of SJ modified silica-gels simultaneously interacts, and passes through chemical bonding, modification
High molecular end-the COOH of ploy (OEGMA) of Silica Surface and peptide molecule-- NH2It is formed and stablizes covalent bond, in turn
There to be chemically active peptide molecule to be fixed on SJ modified silica-gels surface.
4) it assembles
The polypeptide microarrays of fixed 6h must assemble in two days.SJ modified silica-gel thin slices are attached to by gum first
On special reaction column, reaction cavity is covered.Biological incubation reactor can use two-sided biological incubation to react according to demand
Device a, that is, reactor is made of two reaction columns and a reaction cavity or single side biological incubation reactor, that is, one
A reactor is made of a reaction column and a reaction cavity.
5) it preserves
The polypeptide microarrays assembled are needed to vacuumize sealing, are stored in 4 DEG C of refrigerator, spare.
3. being detected with kit
Checking procedure：
1) temperature of reagent is balanced：All reagents are balanced to room temperature.
2) preparation of washing lotion：Washing lotion purified water will be concentrated or distilled water presses 1:9 dilutions, such as：450mL purified waters or
50mL is added in distilled water and concentrates washing lotion, mixes well.The washing lotion being not used is placed on 2~8 DEG C of preservations, can save 3 months.
3) dilution of sample：Sample to be tested serum dilution is pressed 1:100 dilutions are (such as：2 μ L serum are added to 198 μ L
Serum dilution mixes well).Sample detection needs the 200 μ L of sample after diluting.
4) wetted chip：Take out the biochip incubation reaction device needed for detection.1mL washing lotions are added, infiltrate chip surface 3
Minute.Discard the washing lotion for impregnating chip.
5) it is loaded：In the present embodiment, it is loaded with single side biological incubation reactor, it is to be measured by what is diluted using liquid-transfering gun
Sample is added by liquid feeding venthole (any one in two) in single side biological incubation reactor, each single side biological incubation
Reactor adds 200 μ L, with sealing paste closing liquid feeding venthole (two).
6) sample incubation：The single side biological incubation reactor for adding sample is placed in 150rpm on horizontal shaker, is incubated at room temperature
30 minutes.
7) it cleans：Reaction column is taken out from the reaction cavity of single side biological incubation reactor, discards liquid in reactor,
It rinses inside reaction column top chip surface and reaction cavity repeatedly 3 times (about using 12mL washing lotions every time, rinse 1-2 minutes).
8) enzyme labelled antibody is incubated：Reaction column and reaction cavity are reconfigured, clamping.Throw off sealing paste, each single side life
The 200 μ L of goat-anti people IgM solution of horseradish peroxidase-labeled are added in object incubation reaction device.Again liquid feeding is closed with sealing paste
Venthole.It is placed in 150rpm on shaking table, is incubated at room temperature 30 minutes.
9) it cleans again：Reaction cavity is discarded, chip surface is carefully cleaned with washing lotion 3 times.
10) assay chromogenic substrate solution is added：Each chip surface is separately added into 100 μ L assay chromogenic substrate solutions, keeps colour developing liquid energy uniform
Be laid on chip surface.
11) result judgement：It for every a serum, observes by the naked eye, whether each polypeptide in statistics kit
There is response (i.e. the visible royal purple chrominance signal for being better than negative control point of naked eyes), and is retouched according in above-mentioned " diagnostic method " part
The criterion 1 stated is judged.
To 457 parts of clinical serum being collected into, using the testing result of mentioned reagent box and using traditional goldstandard method-
The testing result of nucleic acid detection method (fluorescent quantificationally PCR detecting kit for using Takara) is compared, as a result such as 1 institute of table
Show.The supplier that 457 parts of above-mentioned clinical serum all has the clinical symptoms of hand-foot-and-mouth disease.
The comparison of the kit of 1 embodiment of table and the fluorescent quantificationally PCR detecting kit of Takara
Accuracy rate=(283+144)/457=93.4%
Sensitivity=283/304=93.1%
Specificity=144/153=94.1%
False positive rate=9/153=5.9%
The above result shows that the present invention has good accuracy rate for detecting enterovirus antibodies in human serum (IgM),
Sensitivity and specificity are suitable for the aetology laboratory auxiliary diagnosis of enterovirus, and easy to operate, are suitble to medical treatment at different levels
It is used with disease control department.
More than, more specific description is carried out to the present invention by embodiment, but be not the limit to the technology of the present invention range
It is fixed.
Sequence table
<110>Shenzhen Children's Hospital
<120>It can be used for diagnosing the kit of human enterovirus infection
<160> 10
<170> PatentIn version 3.2
<210> 1
<211> 20
<212>Amino acid
<213>Artificial sequence
<400> 1
YHSSVYSLPPDPDHFDGYKQ 20
<210> 2
<211> 20
<212>Amino acid
<213>Artificial sequence
<400> 2
HPYVLDAGIPISQLTVCPHQ 20
<210> 3
<211> 20
<212>Amino acid
<213>Artificial sequence
<400> 3
ISDLLASVDSEEVRQYCRDQ 20
<210> 4
<211> 20
<212>Amino acid
<213>Artificial sequence
<400> 4
GHFTMLGVRDRLAVLPRHSQ 20
<210> 5
<211> 20
<212>Amino acid
<213>Artificial sequence
<400> 5
AREKVEFLNNLKQLPLLENQ 20
<210> 6
<211> 20
<212>Amino acid
<213>Artificial sequence
<400> 6
GKGELCAVFRADPGRNGPWQ 20
<210> 7
<211> 20
<212>Amino acid
<213>Artificial sequence
<400> 7
EDTHPPYKQTQPGADGFELQ 20
<210> 8
<211> 20
<212>Amino acid
<213>Artificial sequence
<400> 8
SLDFALSLLRRNIRQVQTDQ 20
<210> 9
<211> 20
<212>Amino acid
<213>Artificial sequence
<400> 9
NKEPAVLHSRDPRLEVDFEQ 20
<210> 10
<211> 20
<212>Amino acid
<213>Artificial sequence
<400> 10
GHFTMLGVRDPLAVLPRHSQ 20
Claims (7)
1. a kind of detection kit comprising one or more solid carriers, and be independently connected on the solid carrier
SEQ ID NO：Polypeptide shown in 1~6.
2. detection kit according to claim 1, wherein the SEQ ID NO：Polypeptide shown in 1~6 independently connects
It is connected on same solid carrier.
3. detection kit according to claim 1, wherein the solid carrier is SJ modified silica-gels.
4. detection kit according to claim 1 is used to diagnose human enterovirus infection.
5. a kind of diagnosis subject whether be people's enterovirus infection method, this method includes：
The SEQ ID NO are detected using the detection kit described in any one of Claims 1 to 44：Polypeptide shown in 1~6
Whether there is response to the IgM in the blood sample in subject source；Wherein,
As the SEQ ID NO：When polypeptide shown in 1~6 all has response to the IgM in the blood sample in subject source,
Judgement should subject is a human enterovirus infection；Otherwise, it is determined that the subject is inhuman enterovirus infection.
6. according to the method described in claim 5, wherein, the blood sample is whole blood, blood plasma or serum.
7. according to the method described in claim 5, wherein, the subject is the subject with hand-foot-and-mouth disease clinical symptoms.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710208431.0A CN108663517A (en) | 2017-03-31 | 2017-03-31 | It can be used for diagnosing the kit of human enterovirus infection |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710208431.0A CN108663517A (en) | 2017-03-31 | 2017-03-31 | It can be used for diagnosing the kit of human enterovirus infection |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108663517A true CN108663517A (en) | 2018-10-16 |
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| CN201710208431.0A Pending CN108663517A (en) | 2017-03-31 | 2017-03-31 | It can be used for diagnosing the kit of human enterovirus infection |
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Inventor after: Zhong Shan Inventor after: Ma Dongli Inventor after: Liu Xiaorong Inventor before: Zhong Shan Inventor before: Ma Dongli |
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Application publication date: 20181016 |